Int.J.Curr.Microbiol.App.Sci (2015) 4(9): 991-995
International Journal of Current Microbiology and Applied Sciences
ISSN: 2319-7706 Volume 4 Number 9 (2015) pp. 991-995
http://www.ijcmas.com
Original Research Article
Isolation and Antibiogram of Klebsiella species from
Various Clinical Specimens
Sunilkumar Biradar1* and C. Roopa2
1
Department of Microbiology, Mahadevappa Rampure Medical College,
Gulbarga, Karnataka, India
2
Department of Microbiology, Navodaya Medical College Hospital and Research Centre,
Raichur, Karnataka, India
*Corresponding author
ABSTRACT
Keywords
Klebsiella
pneumoniae,
ESBL
Klebsiella is an important nosocomial human pathogen that has the potential to
cause severe infections. K. pneumoniae is gaining renewed interest because of
emergence of multidrug resistance due to ESBL production. Total 100 samples
showing Klebsiella species on culture were studied. Klebsiella pneumoniae was the
most common species followed by Klebsiella oxytoca. Males were most affected.
Diabetes, alcoholism & previous surgeries were commonly associated factors.
Common infections caused by Klebsiella were wound infections, urinary tract
infections and respiratory infections. Around a quarter of all Klebsiella isolates
were ESBL producers exhibiting multidrug resistance to commonly used
antibiotics like cephalosporins. All isolates were sensitive to carbapenems.
Introduction
virulence factors such as capsular
polysaccharides, lipopolysaccharide and
iron-scavenging systems (siderophores).
Klebsiella species is a Gram negative
opportunistic nosocomial pathogen and is
known to cause community acquired
infections. Klebsiella is known to produce
bacterial pneumonia, urinary tract infection,
wound infections, blood infections and
infections in the intensive care unit
(Podschun and Ullmann, 1998; Brisse et al.,
2006).
Major risk factor for colonisation or
infection with ESBL producing organisms
are long term antibiotic exposure, prolonged
intensive care unit stay, nursing home
residency, severe illness, residence in an
institution with high rates of ceftazidime and
other third generation cephalosporins use
and instrumentation or catheterization
(Sarma et al., 2011). In recent years,
outbreaks of infection caused
by
multidrug-resistant ESBL-KP have been
Nosocomial Klebsiella infections are mainly
caused by Klebsiella pneumoniae followed
by Klebsiella oxytoca (Jadhav et al., 2012).
Klebsiella spp have several
991
Int.J.Curr.Microbiol.App.Sci (2015) 4(9): 991-995
cotrimoxazole
(25μg)
nitrofurantoin
(300μg). Detection of ESBL was done using
CLSI phenotypic confirmation test.
reported throughout the world (Christian et
al., 2010; Filippa et al., 2013).
The study has been conducted to know the
prevalence of Klebsiella species in terms of
samples type, various risk factors and to find
the multidrug resistance including ESBL
producing rates.
Results and Discussion
During the study period, a total of 100 nonrepetitive isolates from various clinical
samples were processed. Among the 100
Klebsiella spp, 89% were Klebsiella
pneumoniae and 11% were Klebsiella
oxytoca. Of the 100 Klebsiella isolates, 62
were from males and 38 were from females
with a male: female ratio of 1.7: 1.
Materials and Methods
Study constituted 100 non-repetitive isolates
from various samples in Microbiology lab of
M.R. Medical College Gulbarga. All the
samples were collected aseptically. Samples
were inoculated on Blood agar and
MacConkeys agar and incubated at 37°C for
24–48 hours. After Gram’s staining,
colonies were subjected to preliminary tests
like catalase, oxidase & to various
biochemical tests like IMViC, TSI, urease,
sugar fermentation tests-glucose, lactose,
sucrose & mannitol. The biochemical
characters used for identification were
positive Voges-Proskauer test, positive
citrate utilization test, positive urease test,
acid and abundant gas production from
glucose, lactose, sucrose, maltose and
mannitol sugar fermentation tests. Antibiotic
sensitivity was done on Mueller-Hinton agar
plates by Kirby-Bauer disc diffusion method
according to the CLSI guidelines (2011). A
log phase broth culture inoculum of the
isolate with a turbidity equivalent to
McFarland 0.5 standard was used. Lawn
cultures on the Mueller-Hinton agar were
prepared and allowed to dry. Antibiotic
discs were applied to the Mueller Hinton
agar surface with the help of sterile forceps.
The antibiotics tested were ampicillin
(10μg), gentamicin (10μg), amikacin (30μg),
amoxicillin/clavulanic
acid
(20/10μg),
piperacillin/tazobactam
(100/10
μg),
cefepime (30μg), cefotaxime (30μg),
ceftriaxone (30μg), ceftazidime (30μg),
ciprofloxacin (5μg), ofloxacin (5μg),
imipenem (10μg), meropenem (10μg),
The highest percentage of Klebsiella spp
were isolated from pus (50%) followed by
urine (21%), sputum (18%), blood (7%),
throat swab (3%) and CSF (1%).
Various risk factors associated with
Klebsiella infection observed were diabetes
mellitus, hypertension, alcoholism, smoking,
post-operative state and, catheterization.
The isolates were 100% sensitive to
imipenem & meropenem, cefepime (82%),
piperacillin/tazobactum (77%), amikacin
(73%), ceftriaxone (69%), cefotaxime
(67%), nitrofurantoin (66%), ceftazidime
(63%), amox-clav (62%), gentamicin (62%),
ofloxacin (59%), ciprofloxacin (56%),
cotrimoxazole (33%) and ampicillin (06%).
Of 100 isolates of Klebsiella, 24 isolates
were found to produce extended spectrum
beta lactamases detected by CLSI
phenotypic confirmation test.
Klebsiella pneumoniae (KP) is one of the
leading causes of nosocomial infections seen
worldwide, causing pneumonia, bloodstream
infections, urinary tract infections, surgical
site infections and meningitis (Peleg and
Hooper, 2010). In the present study 100
isolates of Klebsiella were studied. In the
current study, isolation rate of K.
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Int.J.Curr.Microbiol.App.Sci (2015) 4(9): 991-995
pneumoniae was more compared to K.
oxytoca. Klebsiella pneumoniae incidence
was 89%. This finding is comparable to the
report of Asmaa (2012).
Males (62%) were more affected than
females, and this could be due to higher
prevalence of alcoholism and smoking in
males.
Table.1 Percentage of Klebsiella species isolated
Species
Klebsiella pneumoniae
Klebsiella oxytoca
No of isolates
89
11
Percentage
89%
11%
Table.2 Percentage of isolation of Klebsiella species from different clinical specimens
Sample
Pus
Urine
Sputum
Blood
Throat swab
CSF
No. of isolates
50
21
18
7
3
1
percentage
50%
21%
18%
7%
3%
1%
Table.3 Various Risk factors in patients with Klebsiella infection
Risk factors
Diabetes
Hypertension
Post-surgical
Catheterisation
Alcohol & smoking
No of patients
36
16
27
08
21
Table.4 Antibiotic sensitivity profile
Antibiotic
Ampicillin
Gentamicin
Amikacin
Amox-clav
Piperacillin/Tazobactum
Cefepime
Cefotaxime
Ceftriaxone
Ceftazidime
Ciprofloxacin
Ofloxacin
Imipenem
Meropenem
Cotrimoxazole
Nitrofurantoin
993
Sensitivity
06%
62%
73%
62%
77%
82%
67%
69%
63%
56%
59%
100%
100%
33%
66%
Int.J.Curr.Microbiol.App.Sci (2015) 4(9): 991-995
The highest percentage of Klebsiella spp
were isolated from pus (50%) followed by
urine (21%), sputum (18%), blood (7%),
throat swab (3%) and CSF (1%). Various
studies done on Klebsiella found similar
results, but most common sample being
urine in their study (Amit Kumar Singh,
2015). Most common risk factor associated
with Klebsiella infections was found to be
diabetes followed by alcoholism and
previous surgeries. Several studies done
suggest these as the most common risk
factors in patients (Namratha et al., 2015;
Abhilash et al., 2010).
Klebsiella oxytoca. Klebsiella were still
100% sensitive to carbapenems, but
sensitivity to cephalosporins ranged from
50–70%. Klebsiella are now being
recognized as one of the major threats to
effective management of patients in hospital,
especially in a developing country like
India.
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